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MB Sample ID: SA093260
Local Sample ID: | S0019_POS |
Subject ID: | SU001349 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Male and female |
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Subject:
Subject ID: | SU001349 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Male and female |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
S0019_POS | SA093260 | FL013365 | PXE | Treatment1 |
Collection:
Collection ID: | CO001343 |
Collection Summary: | All materials were collected from human donors without identifiers under institutional review board exemption/approval. We acquired 31 aqueous humor (AH) samples from participants at the Veterans Administration (VA) Medical Center (Miami, FL). Patient samples used for Isotopic Ratio Outlier Analysis (IROA) consisted of individuals with PEX (n=31). |
Sample Type: | Eye tissue |
Collection Location: | 1201 NW 16th St, Miami, FL 33125 |
Treatment:
Treatment ID: | TR001364 |
Treatment Summary: | No Treatment |
Sample Preparation:
Sampleprep ID: | SP001357 |
Sampleprep Summary: | The 31 samples were prepared for isotopic ratio outlier analysis (IROA). Approximately 800 µL of precipitate solution (8:1:1 Acetonitrile: Methanol: Acetone) was added to the 50 µL of aqueous humor. The metabolites were vortexed and incubated at 4 °C for 30 minutes. This was followed by another incubation period at -20 °C for 1 hour. Each sample was centrifuged at 20,000 x g for 10 minutes at 4 °C (Beckman Microfuge 18) to form a pellet and 375 µL of supernatant was collected. The supernatant was dried in a speed vacuum for approximately 20 minutes or until the sample was fully dry and stored at -20 °C. The IROA internal standard (IROA-IS, IROA technologies) (U-95% 13C) was reconstituted in 1.2 mL of LC-MS grade water. The samples were reconstituted in 25 µL of LC-MS grade water and 10 µL was combined with 20 µL of IROA-IS and subjected to LC-MS/MS analysis. A long-term reference standard (LTRS) was reconstituted in 50 µL of LC-MS grade water and subjected to LC-MS/MS as the first, last, and every 10 samples. |
Combined analysis:
Analysis ID | AN002118 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Thermo Accela 600 |
Column | ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap |
Ion Mode | UNSPECIFIED |
Units | C12:C13 Ratio |
Chromatography:
Chromatography ID: | CH001551 |
Instrument Name: | Thermo Accela 600 |
Column Name: | ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) |
Column Temperature: | 40 C |
Flow Rate: | 350 ul/min |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 100% acetonitrile; 0.1% formic acid |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS001973 |
Analysis ID: | AN002118 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Samples were ran in positive and negative ion mode Data dependent MS/MS |
Ion Mode: | UNSPECIFIED |